1. Field of Invention
The invention relates to a method for treatment of leukemia in mammals, and more specifically chronic myeloid leukemia (CML), using reactive oxygen intermediates. Reactive oxygen intermediates are administered in a therapeutically effective amount to a mammal that has leukemia. The administration of reactive oxygen intermediates, and more specifically ozone, has been found to be particularly effective in the treatment of CML and on the modulation of the immune and hematopoietic systems of mammals having cancer.
2. Description of Prior Art
Therapeutic usage of ozone has occurred in Europe for more than 50 years. The lack of published data regarding controlled trials as well as mechanisms of explaining it""s actions have made ozone therapies almost completely unknown in North America.
Ozone, reactive oxygen intermediates (ROI) or reactive oxygen species (ROS) are known to damage cell membranes and cell proteins. More specifically, in respect of cell proteins, ozone or ROI""s exert deleterious effects on amino acids namely, histidine and tyrosine. The mechanisms behind this protein damage have not been fully elucidated. However, it is believed to be related to the chemical reaction between the reactive oxygen radicals hydroxyl radical, hydrogen peroxide, super oxide anion, (O2xe2x88x92, H2O2 , and OH), (hereafter referred to as ROI) and the chemical bonds in the proteins or amino acids. Most healthy normal cells have anti-oxidant enzymes such as catalase, peroxidase, superoxide dismutase and glutathione, which function to remove or neutralize these oxygen radicals so that no damage occurs. However, it is theorized that ozone and ROI""s may cause damage by removing or damaging tyrosine contained in proteins, or by signalling the cell to stop production of protein tyrosine kinases or inhibit the protein tyrosine kinase function which is important for signal transduction as it leads to cell proliferation in normal and transformed cells.
Ozone and ROI""s are also known to induce oxidation of cell membrane molecules and have been previously shown to have antineoplastic properties (Viebahn, 1994), more specifically the ability to synergize with chemotherapeutic agents (Zanker, 1988; Zanker and Kroczek, 1990) and radiation (Karlic, 1987). However, the use of ozone and ROI""s as an agent in the treatment of cancer has been limited.
Current research suggests that ozone achieves its antineoplastic effects by inducing cell death and inhibiting cell growth in cancerous cells. However, the literature does not indicate the method of cell death. Ozone or ROI induced cell death may be caused by necrosis (typical rupture of cellular membranes), lipid peroxidation, or signalling the cell to commit suicide (called programmed cell death or apoptosis).
In one example, Sweet et al. (1980) examined the effect of ozone on solid tumour masses. The results of these studies indicated ozone can selectively inhibit the growth of cultured human cancerous cell lines derived from lung, breast and uterine solid tumours with little damage to normal cell growth.
Alternative medicine has also referred to the use of ozone or oxygen therapy in reference to cancer and specifically acute T-cell lymphoma (Sartori, 1994). The results of this study demonstrated that ozone in combination with other therapies appeared to have some beneficial effects on the lymphomas and assorted cancers. In general, approximately 50% of the patients died as a result of their aliments. In the surviving patients, there was no clear documentation that ozone or ROI""s were the therapeutic agent acting on the cancerous lymphoma. However, Sartori""s experiments were concerned with the treatment of 12 lymphoma cases and several of the patients had other complications including AIDS and other cancers. Indeed, clear conclusions were difficult to draw from the Sartori cases as they were not part of a proper control clinical trial and several of the patents also received chemotherapy. Furthermore, the Sartori patients received a variety of alternative medicine treatments called the Life Science Universal (L.S.U.) in combination with ozone.
Neither of these researchers examined the effect of ozone or ROI""s on any of the leukemias including chronic myeloid leukemia, nor did they address the use of ozone and ROI""s to differentiated cancerous cells as a method of stopping cancer. Indeed, it is well known in cancer research, that the results obtained with one anti-cancer agent in the treatment of one type of cancer, rarely has similar effects in other types of cancer.
Current research has also theorized about the effects ozone has on cytokine levels or cytokine profile. In this respect there is a network or control function for the hematopoietic system which is influenced by a variety of negative and positive stimuli (Gregory et al, 1991; Ogawa, 1993; Kuby, 1994). Negative regulators include interferon alpha, beta, and gamma (IFN-xcex1, IFN-xcex2 and IFN-xcex3), tumour necrosis factors alpha and beta (TNF-xcex1 and xcex2) and prostaglandins. Positive regulators of hematopoiesis include the colony stimulating factors (CSF) such a granulocyte/monocyte (GM-CSF), granulocyte (G-SF), monocyte (M-CSF), interleukin-3 (IL-3 or multi-CSF) and stem cell factor (SCF). In addition to the CSFs, other positive regulators of hematopoiesis include IL-1, IL-4, IL-5, IL-6 and IL-11 (Gregory et al, 1991; Ogawa, 1993, Kuby, 1994).
Several studies have shown that ozone increases cytokine levels such as IL-1, IL-2 and IFN-xcex3 in humans (Bocci, 1990; Bocci et al., 1993; Bocci, 1994, Bocci, 1995). IL-2 also activates the release of (IFN-xcex3) and other regulators which are known to activate the hematopoietic and immune systems (Kuby, 1994; Cruse and Lewis, 1995). None of these studies have examined the effects of ROI""s on the immune and hematopoietic systems with respect to cancer and the leukemias.
Another aspect of the abnormalities in leukemia and more specifically CML, is the lack of cell maturation, including cellular enzyme systems and their control mechanisms (Clarkson and Strife, 1993). More specifically, an o-alkyl cleavage enzyme which normally metabolizes alkyl-lysophospholipids or ether lipids is altered or inhibited in leukemia. The accumulation of these alkyl-lysophospholipids such as ET-18-OCH3, interferes with normal phospholipid metabolism and membrane composition resulting in damaged membranes. Normal cells are not adversely affected (Andreesen et al., 1979) by ether lipids as the o-alkyl cleavage enzyme catabolizes the lipids. Previous studies on ether lipids in leukemias did not examine the synergism between ozone or ROI therapy in conjunction with alkyl-lysophospholipids or ether lipids.
Other cancer therapies have focused on bone marrow transplants, but despite improvement of allogenic bone marrow transplant technologies as a means to combat the leukemias, no therapies are currently available for patients who lack a bone marrow donor. Typical maintenance agents such as interferon and hydroxyurea have shown some survival advantage but these therapies are by no means curative. In addition, the toxic side effects of xe2x80x9cconventionalxe2x80x9d chemotherapeutic agents significantly decreases the quality of life, while not necessarily extending it. Once the patient enters the terminal phase of the disease, called blast crisis, there are no treatment options available. It is at this stage that any therapy that provides an extension of life or improves the quality of life is greatly needed.
It would therefore be desirable to provide a method of treatment for leukemia which overcomes the deficiencies of the prior art.
It is an object of the present invention to overcome the deficiencies in the prior art treatments of mammals suffering from leukemia such as CML. In accordance with one aspect of the present invention, there is provided a therapy which results in surprising antineoplastic effects in the treatment of leukemia such as CML. More specifically the invention is the administration of a therapeutically effective amount of O3 to the blood of a mammal having leukemia. Preferably, the invention provides treatment for leukemia such as CML, by administering a therapeutically effective amount of ozone so as to induce cellular differentiation of the leukemic cells.
In accordance with a second aspect of the invention there is provided a method of treating leukemia in mammals by administering therapeutically effective amounts of O3 and ether lipids. Preferably, the invention includes the administration of ozone and alkyl-lyophospholipids in therapeutically effective amounts to the blood of mammals having leukemia so as to induce cellular differentiation of the leukemic cells.
In accordance with a third aspect of the invention there is provided a method of treating cancer by administering therapeutically effective concentrations of O3 thereby targeting and/or modulating the hematopoietic and immune systems by the activation of or inhibition of these systems. Preferably, the invention provides that ozone may alter the hematopoietic system through selective targeting and modulating the immune system to combat cancers and leukemias.
In accordance with a fourth aspect of the invention there is provided a method of generating ozone by an ozone generator and administering a therapeutically effective concentration of reactive oxygen intermediates for a period of 5-20 minutes.
With respect to all aspects of the invention the modes of administering reactive oxygen intermediates include but are not limited to: 1. direct injection of gas containing ozone and ROI, 2. ex vivo treatment of blood with ozone and ROI""s followed by reinfusion of treated blood, 3. injection of ozonated products, 4. inhalation of ozonated products, 5. insufflation with ozone or ROI gas.
Finally, the invention provides for the generation of ozone and the use of ozone and ROI therapy in a clinical setting to treat mammals having leukemia. Another aspect of the invention is for the use of reactive oxygen intermediates for the production of a medicament for the treatment of leukemia. The administration of ozone and ROI may result in the induction of cell differentiation and maturation of the leukemic blast cells that remain after treatment. This would prove beneficial to the clinical patient by: 1. Reducing the leukemic cell burden on the patent""s hematopoietic systems (bone marrow, spleen, and liver) as well as the peripheral blood circulation; 2. Stimulating the immature blast cells to mature and differentiate into normal blood cells; 3. Allowing the differentiation and proliferation of the normal hematopoietic stem cells; and 4. Increasing the psychological and physiological well being of the patient.